Grant: Research Tools and Instruments - Category 1 (<$150,000)

Confocal Laser Scanning Microscopy (CLSM) is a powerful tool that enables biologists to obtain optical laser 'slices' through solid tissue or cells without the need for cutting physical sections. Three-dimensional reconstructions of whole cells/tissues can be assembled from these optical sections. Molecules of interest in tissues or single cells can be identified using antibodies to them and fluorescent tags that glow when illuminated by particular wavelengths of laser light. CLSM is thus a central, powerful tool for very many cell biologists and physiologists worldwide. The York Biology Dept CLSM facility was installed in 2002. CLSM equipment and techniques have improved greatly since then. About 10000hr/year are logged on our CLSM by a large segment of the department, and it is in desperate need of some new components and replacement of certain old ones. It has only red and green lasers. It is important that we obtain a 405nm (blue) laser. This will enable visualization of three different molecules simultaneously and permit analysis of the relationships of several cellular structures with each other. It will also enable use of stains on living cells (eg DAPI), permitting time-lapse studies of movements of subcellular structures, with minimal photobleaching by the laser. Many fluorescent staining procedures require several days at present to complete; we wish to acquire a novel microwave system that accelerates penetration of molecules through membranes without heating them, which will reduce processing time to a few hours and allow researchers to achieve much more in the same time. Finally, several original components of the 2002 CLSM have reached the end of their useful life and require replacement if the CLSM is to continue to operate. Even the computer is 8 years old and cannot run the current software required by the CLSM manufacturer. York University has a distinguished research record in both cell biology and physiology, but this can be maintained only if the requested enhancements are installed in our departmental CLSM.